biostudies-literatureYang KNational Institutes of HealthNIGMS NIH HHS100687https://www.ebi.ac.uk/biostudies/studies/S-EPMC8348308biostudies-literatureUnknown2(3)Carnitine palmitoyltransferase-1 (CPT-1) is a rate-controlling enzyme for long-chain fatty acid oxidation. This manuscript provides protocols for measuring CPT-1-mediated respiration in permeabilized, adherent cell monolayers and mitochondria freshly isolated from tissue, along with examples to assess the potency and specificity of interventions targeting CPT-1. Strengths of the approach include ease, speed, and breadth of analysis, whereas drawbacks include loss of physiological regulation in reductionist systems and indirect assessment of CPT-1 enzymatic activity. For complete details on the use and execution of this protocol, please refer to Divakaruni et al. (2018).STAR protocolsMeasuring CPT-1-mediated respiration in permeabilized cells and isolated mitochondria.PMC8348308R35GM138003P30DK063491R35 GM138003Doan MTYang KStiles LDivakaruni ASfalseMeasuring CPT-1-mediated respiration in permeabilized cells and isolated mitochondria.Carnitine palmitoyltransferase-1 (CPT-1) is a rate-controlling enzyme for long-chain fatty acid oxidation. This manuscript provides protocols for measuring CPT-1-mediated respiration in permeabilized, adherent cell monolayers and mitochondria freshly isolated from tissue, along with examples to assess the potency and specificity of interventions targeting CPT-1. Strengths of the approach include ease, speed, and breadth of analysis, whereas drawbacks include loss of physiological regulation in reductionist systems and indirect assessment of CPT-1 enzymatic activity. For complete details on the use and execution of this protocol, please refer to Divakaruni et al. (2018).2021-01-01T00:00:00Z2021 Sep2025-04-05T08:50:24.948Z2025-02-19T01:15:59.243ZS-EPMC83483083440177310.1016/j.xpro.2021.100687